UHPLC-MS分析人促红素产品中CHO宿主细胞蛋白残留  

Analysis of residual CHO host cell protein in human erythropoietin products by UHPLC-MS

作　　者：于雷[1] 秦玺[1] 张晓夕 史新昌[1] 李响[1] 周勇[1] YU Lei;QIN Xi;ZHANG Xiao-xi;SHI Xin-chang;LI Xiang;ZHOU Yong(Key Laboratory of the Ministry of health for Research on Quality and standardization of Biotech Products,National Institute of Food and Drug Control,Beijing 100050,China;Thermo Fisher Scientific(China)Co.,Ltd.,Shanghai 201210,China)

机构地区：[1]中国食品药品检定研究院,卫生部生物技术产品检定方法及其标准化重点实验室,北京100050 [2]赛默飞世尔科技(中国)有限公司,上海201210

出　　处：《药物分析杂志》2023年第8期1402-1407,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：国家质量基础设施体系(2021YFF0600804)。

摘　　要：目的:利用超高效液相色谱-质谱联用技术(UHPLC-MS)分析人促红素(human erythropoietin,hEPO)产品中CHO宿主细胞蛋白(host cell protein,HCP)残留。方法:将hEPO原液和蛋白标准品一起进行变性、还原烷基化、酶切处理;采用Accucore Vanquish C18色谱柱串联超高效液相色谱和Q ExactiveTM Plus组合型四极杆OrbitrapTM质谱仪对样品进行分离和检测;使用BioPharma Finder 4.1软件中的HCP工作流程进行定性及定量分析。结果:10种HCP在4批hEPO原液样品中均被检测到,且不同样品中HCP总量有较大差异,从25×10^(-6)至约1000×10^(-6);含量>50×10^(-6)的HCP,CV值均<10%。结论:UHPLC-MS法可对CHO细胞表达的hEPO产品中HCP进行定性定量分析,不同来源hEPO样品中HCP种类和含量均有差异。Objective:To analyze residual CHO host cell protein(HCP)in human erythropoietin(hEPO)prod⁃ucts by UHPLC-MS.Methods:The hEPO bulks and internal protein standard were denatured,reduced,alky⁃lated and digested;Accucore Vanquish C18 column connected with UHPLC and Q Exactive Plus Hybrid Quadru⁃pole-Orbitrap Mass Spectrometer were employed to separate and detect the samples;qualitation and quantitation of HCP were performed using HCP workflow in BioPharma Finder 4.1 software.Results:Ten HCPs were detected in all 4 batches of hEPO bulks,and the HCP amounts varied between different samples,ranging from 25×10^(-6)to approximately 1000×10^(-6);the CV%were below 10%for the HCPs above 50×10^(-6).Conclusions:The meth⁃od of UHPLC-MS could be used for qualitative and quantitative analysis of HCP in hEPO products expressed in CHO system,and the HCP types and amounts in different samples varied considerably.

关 键 词：人促红素 CHO细胞表达系统 宿主细胞蛋白 超高效液相色谱-质谱联用技术 中国仓鼠全蛋白库 

分 类 号：R917[医药卫生—药物分析学]